“Nuclease-free” Gene Editing for Sickle Cell Disease and β-Thalassemia
Faculty Information
Name:
Prof. Arun Srivastava
Email
aruns@ufl.edu
Phone
(352) 273-8259
Faculty Department/Division
Pediatrics
This project is primarily:
Basic
Research Project Description:
Human hemoglobinopathies (sickle cell disease and -thalassemia), are by far the most common (1 in 600) monogenic diseases worldwide. Although gene editing with several nucleases (mega TAL, zinc-finger nuclease, CRISPR/Cas9, base editors, prime editors) have been performed, and remarkable efficacy has been achieved in animal models, the use of these nucleases in humans has not been optimal. For example, serious adverse events, such as immune responses to these bacteria-derived nucleases, off-target cutting of DNA, chromosomal translocations, and loss of chromosomes, and genotoxicity have been observed. The lack of specificity of vehicles, such as lipid nanoparticles (LNPs), to deliver these nucleases in vivo also remains a challenge. Thus, there is need for the development alternative strategies for gene editing that are safe and efficient, as well as the development of a highly efficient and specific delivery vehicle. We have developed strategies to overcome both of these limitations. For example, we have developed a decoy zinc-finger transcription factor synthetic peptide capable of gene editing without causing double-stranded breaks in DNA. We have also identified adeno-associated virus 6 (AAV6) for high-efficiency transduction of primary human hematopoietic stem cells (HSCs). We have generated next-generation AAV6-human parvovirus B19 (B19V) hybrid and AAV-B19V chimeric vectors for high-efficiency transduction and erythroid lineage-restricted expression from B19V promoter of synthetic decoy zinc finger transcription factor peptides for “nuclease-free” gene editing for human sickle cell disease and -thalassemia. Medical students will be involved in evaluating the efficacy of further optimized AAV6-B19V vectors to achieve safe and efficacious gene editing in a xenograft mouse model. Funding for the project is provided by the National Institutes of Health and the George H. Kitzman Endowment.
Song, M.A. Kauss, E. Kopin, M. Chandra, T. Ul-Hasan, E. Miller, G.R. Jayandharan, A.E. Rivers, G.V. Aslanidi, C. Ling, B. Li, W. Ma, X. Li, L.M. Andino, L. Zhong, A.F. Tarantal, M.C. Yoder, K.K. Wong, Jr., M. Tan, S. Chatterjee, and A. Srivastava. Optimizing the transduction efficiency of capsid-modified AAV6 vectors in primary human hematopoietic stem cells in vitro and in a xenograft mouse model in vivo. Cytotherapy, 15: 986-998, 2013.
Song, X. Li, G.R. Jayandharan, Y. Wang, G.V. Aslanidi, C. Ling, L. Zhong, G. Gao, M.C. Yoder, C. Ling, M. Tan, and A. Srivastava. High-efficiency transduction of primary human hematopoietic stem cells and erythroid lineage-restricted expression by optimized AAV6 serotype vectors in vitro and in a murine xenograft model in vivo. PLoS One, 8(3): e58757, 2013.
Ling, K. Bhukhai, Z. Yin, M.Q. Tan, M.C. Yoder, P. Leboulch, E. Payen, and A. Srivastava. High-efficiency transduction of primary human CD34+ hematopoietic stem/progenitor cells by AAV6 serotype vectors: Strategies for overcoming donor-variation and implications in genome editing. Scientific Reports, 6, 35495, 2016.
Yang, K. Qing, G.D. Keeler, L. Yin, M. Mietzsch, C. Ling, B.E. Hoffman, M. Agbandje-McKenna, M. Tan, W. Wang, and A. Srivastava. Enhanced transduction of human hematopoietic stem cells by AAV6 vectors: Implications in gene therapy and genome editing. Molecular Therapy-Nucleic Acids. 20: 451-458, 2020.
Does this project have an international component or travel?
No
Neurological Complications in Fontan Patients Undergoing Cardiac Transplantation
Name: Wei Wang, MD
Email: wwang29@peds.ufl.edu
Division: Pediatric Cardiac Critical Care Medicine, Cardiology, Neurology, Pediatrics
Background:
Patients with Fontan circulation are at risk for neurological complications due to a prothrombic state, chronic microemboli, and multiple previous palliations. We describe neurological complications in patients with Fontan circulation in the early perioperative period after cardiac transplantation.
Methods:
This will be an extension of a previous retrospective, single-center case control study which evaluated the post- heart transplant neurological outcomes in all patients with Fontan physiology (4-53 years) who underwent cardiac transplantation at University of Florida. Three points of evaluation were used: (1) neurological findings on CT and MRI in the first month following transplant, (2) abnormalities on electroencephalography (EEG), and (3) the initiation of antiepileptic medications prior to post-transplant discharge. Data collected will include the patients’ preoperative anticoagulation, inotropic support, end organ function, and hemodynamics.
Role of the medical student:
Drafting and submitting a new IRB for the extension of the previous study
Making a redcap data base
Reviewing patients charts and inputting pertinent information into the RedCap database
Opportunities for poster and abstract presentations available
additional opportunities for shadowing available within the pediatric cardiac critical care unit and congenital heart center operating rooms
Med student should ideally have a strong interest in pediatric or adult cardiology or neurology. Student will work closely with Dr. Tavenner Dibert, Dr. Wei Wang and Dr. Joseph Philip from the division of pediatric cardiac critical care medicine. Students should be proactive, timely in email/text communication, and have strong communication skills. Schedule is flexible and a portion of this work may be able to be completed remotely. However, in person meetings may be required for EPIC training, patient chart review, redcap database set up etc.
Does this project have an international component or travel?
No